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DISP3/PTCHD2 function in neural cells
Konířová, Jana ; Bartůněk, Petr (advisor) ; Anděrová, Miroslava (referee) ; Pacherník, Jiří (referee)
DISP3 protein, also known as PTCHD2, belongs to the PTCHD family of proteins, which contain a sterol-sensing domain in their structure. The expression of the Disp3 gene is high in neural tissues and is regulated by thyroid hormone. The DISP3 gene is associated with development and progression of certain types of tumors, as well as with development of some neural pathologies. Neural stem cells also display high expression of the Disp3 gene. Neural stem cells are defined by their capability to self-renewal and capacity to differentiate into the basic types of neural cells - neurons, astrocytes, and oligodendrocytes. Precise regulation of the balance between proliferation and differentiation of neural stem cells is crucial for development of the central nervous system and its subsequent proper functioning, and disruption of this balance may lead to development of various pathologies. In this work we mainly focused on describing the function of the DISP3 protein in neural cells and tissues. We have shown that during differentiation of neural stem cells, the expression of the Disp3 gene is significant decreased. Furthermore, we have found that in neural stem and progenitor cells, the increased expression of the Disp3 gene promotes their proliferation. Moreover, when Disp3 expression was disrupted, the...
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DISP3/PTCHD2 function in neural cells
Konířová, Jana ; Bartůněk, Petr (advisor) ; Anděrová, Miroslava (referee) ; Pacherník, Jiří (referee)
DISP3 protein, also known as PTCHD2, belongs to the PTCHD family of proteins, which contain a sterol-sensing domain in their structure. The expression of the Disp3 gene is high in neural tissues and is regulated by thyroid hormone. The DISP3 gene is associated with development and progression of certain types of tumors, as well as with development of some neural pathologies. Neural stem cells also display high expression of the Disp3 gene. Neural stem cells are defined by their capability to self-renewal and capacity to differentiate into the basic types of neural cells - neurons, astrocytes, and oligodendrocytes. Precise regulation of the balance between proliferation and differentiation of neural stem cells is crucial for development of the central nervous system and its subsequent proper functioning, and disruption of this balance may lead to development of various pathologies. In this work we mainly focused on describing the function of the DISP3 protein in neural cells and tissues. We have shown that during differentiation of neural stem cells, the expression of the Disp3 gene is significant decreased. Furthermore, we have found that in neural stem and progenitor cells, the increased expression of the Disp3 gene promotes their proliferation. Moreover, when Disp3 expression was disrupted, the...
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Transcriptional regulation of GAL genes - model for chromatin structure study
Brožková, Anita ; Folk, Petr (advisor) ; Konířová, Jana (referee)
The GAL system in Saccharomyces cerevisiae enables this budding yeast to metabolize galactose. Expression of GAL genes is controlled by a regulatory cascade in which galactose triggers the activation of GAL gene expression, whereas glucose acts as a repressor. GAL genes in yeast have been used for decades as a model system for transcription regulation in eucaryotes. The products of GAL genes are GAL regulatory and GAL structural genes. Intriguingly, during studies of the GAL system it has been discovered that one of the regulatory genes and the structural gene for the galactokinase enzyme are apparently related. It has been suggested that an ancestor of the two genes underwent a gene duplication event which allowed the paralogs to gain different functions. The GAL genes serve as a model system for the study of chromatin changes during transcription activation or repression. Transcriptional repression of GAL1 and GAL10 genes via ncRNA represents one of the recently discovered regulatory mechanisms of the GAL system. This mechanism has been discovered due to the changes in the histone methylation pattern across the GAL10 locus. However, the latest discovery in GAL gene regulation has probably been the role of nuclear localization of the GAL1,-7,-10 cluster in response to changes of carbon sources. GAL genes...
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The TUB3 intron splicing in PRP45 mutant cells
Konířová, Jana ; Půta, František (advisor) ; Zimmermannová, Olga (referee)
Protein Prp45, an essential factor of the yeast Saccharomyces cerevisiae, is implicated in pre-mRNA splicing. A truncated version of the PRP45 gene, prp45(1-169), which exhibits a temperature sensitivity, was previously prepared in our laboratory. The aim of this work is to contribute to better understanding of prp45(1-169) mutant phenotype. We tested the prp45(1-169) strain for its response to microtubule inhibitor benomyl and then we found that TUB3 overexpression from plasmid rescues discovered prp45(1-169) mutant cells hypersensitivity to benomyl. In addition, we studied the influence of TUB1, TUB3, and COF1 intron deletion on prp45(1-169) strain temperature sensitivity. Using RT-qPCR method we found that prp45(1-169) mutation results in the distinctive increase of pre-mRNA level for all tested genes, that could implicate that pre-mRNA splicing in these cells is affected before first transesterication.
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